DE69928000T2 - BREAST CANCER TREATMENT BY NATURAL ESSENTIAL OILS - Google Patents

Description

Territory of invention

The The present invention relates generally to use of compounds of essential vegetable oils for the production of drugs for prophylactic or therapeutic treatment of breast cancers in mammals, including People.

background the invention

breast cancer is a proliferative disease of mammary epithelial cells and it has been shown that estrogen the Cell proliferation of these cells stimulated both in culture and in mice (Soto and Sonnenschein 1985; Osborne, 1988). It has been suggested that xenoestrogens cell proliferation by binding and activating estrogen receptors (ERs) (Miller et al., 1993, Hoffman, 1992). The appearance of breast cancer took while the past two or three decades steadily increasing, a trend that characterized by increasing rates among estrogen sensitive Tumors, by continued increases among older women and by growing Numbers in both industrialized countries as well as in developing countries (Harris et al., 1992). Between 1973 and 1980, the occurrence increased breast cancer in the United States by a small 8% among women age under 50, while it is 32.1% among women in the age group of 50 years or older (Ries et al., 1991). This upward shift is consistent with the historical pattern of accumulation of Organochloride insecticide residues (xenoestrogens) in the environment (Mussalo-Rauhamaa et al., 1990; Wolff et al., 1993; Davis et al., 1993). Breast cancer is also the second leading cause Cancer deaths in women and it is estimated that there were additional in 1998 43,900 deaths due to breast cancer will occur. It has been suggested that environmental estrogens or endocrine disruptors play a role in the ethology or promotion of breast cancer (Davis et al., 1993; Dewailly et al., 1995). Experimental evidence shows that xenoestrogens are the source of estrogen production and metabolism and are among the risk factors, causing breast cancer (Nelson, 1994; Berthois et al., 1986; Henderson et al., 1993; Jobling et al., 1996; Dees et al., 1997). Most of the known risk factors for breast cancer that are at least 30% of the cases contribute (Henderson et al., 1993), with overall lifetime exposure to reproductive Chemicals, like estrogen and xenoestrogens connected.

It It appears plausible that soft tissue cancer in mammals is a result of increasing estrogen levels and increasing exposure Environmental xenoestrogens increases every year. For example, the number of prescriptions increases of estrogen for women in menopause fast and is currently on 50 million prescriptions yearly valued in the United States alone. This rising consumption of estrogen partly contributes to the higher Risk of breast cancer in both young women and women in at middle age. estrogen is in all mammals present and it is in women for reproductive organs such as the ovaries, uterus, breast, etc. essential. In men is however estrogen for the Sperm production and maturation required. The abusive Use of estrogen, that for Women is at least partially committed to development soft tissue cancers, especially breast cancer. It is therefore desirable the adverse effects of estrogen to fight in women or to counteract them.

The current, FDA-approved treatments in the United States, e.g. Tamoxifen, to some extent, are in a part of the female population while fighting the adverse effects of estrogen effective. Unfortunately These treatments are not completely effective and can be self-directed additional cause health related effects, such as uterine cancer. Therefore it is, if one could identify connections, the the present Make treatments more effective or in connection with or instead of existing treatments could act possible that some of these adverse side effects are alleviated or even eliminated would. A possible The source of alternative treatments are natural, non-toxic compounds. It is suggested that these compounds be advantageous for safer and more effective treatments.

The Use of certain ethereal Monoterpenoid vegetable oils (Alpha-terpineol, Linalool and limonene) is considered a potential treatment for breast cancer proposed. However, these monoterpenoids are not completely effective and have proven to be weak anti-proliferative cancer products. additionally do not put this data the ability These compounds suggest to combat the effects of estrogen. This Can raise the question of how this product in women with an estrogen drug interact can.

Accordingly, there it a big one Need for new pharmaceutical compositions containing non-toxic ingredients that are effective in preventing or treating Soft tissue cancer in mammals can be used.

Sylvie Bardon et al. Report in "Nutrition and Cancer ", volume 32, No. 1, 1998, pages 1-7, that monoterpene cell growth, cell cycle progression and cyclin D1 gene in human Inhibit breast cancer cell lines.

FR-A-2 706 771 describes a pharmaceutical composition for treating of cancer, comprising at least one compound of an ethereal Vegetable oil like eugenol, limonene or thymol.

DE 38 29 200 A discloses pharmaceutical compositions comprising eugenol and isoeugenol for treating the immunodeficiency disease AIDS.

EP 0 264 660 A discloses a pharmaceutical composition comprising thymol or carvacrol for use as a dental varnish or cement.

WHERE 93/11775 A discloses a pharmaceutical composition comprising Thymol, carvacrol, citral etc., as a penetration enhancing agent Compound in transdermal patches.

DE 35 15 253 C discloses pharmaceutical compositions comprising cinnamic alcohol for biological cell and tissue regeneration.

FR 2 697 133 A discloses pharmaceutical compositions comprising eugenol, isoeugenol, cinnamaldehyde, etc. for biocidal or biostatic uses.

WHERE 00/25802 A discloses an anti-tumor composition, the benzyl alcohol includes. This document is state of the art according to Art. 54 (3) EPC.

WHERE 99/48469 A discloses a pharmaceutical composition comprising Menthol, eucalyptus oil, Thymol, citronella oil or limes. This document is state of the art according to Art. 54 (3) EPC.

WHERE 99/48386 A discloses a pharmaceutical composition comprising D-limonene as an anticancer drug. This document is stand the technique according to Art. 54 (3) EPC.

WHERE 00/24409 A discloses a pharmaceutical composition comprising Eugenol, cineole and cinnamaldehyde. This document is state of the art according to Art. 54 (3) EPC.

Summary the invention

The The present invention provides the new use of certain essentials Vegetable oils, of course or synthetic in its source, or mixtures or derivatives thereof for the manufacture of a medicament as prophylactic for or a Treatment of breast cancer.

The Above and other tasks are covered by the present Invention solved, the use of eugenol, thymol, isoeugenol, benzyl alcohol, Carvacrol, cinnamyl alcohol, cinnamic aldehyde, citronellal or trans-anethole for the manufacture of a medicament for the treatment of breast cancer is directed. Also here are pharmaceutical compositions described that at least one compound of an ethereal Vegetable oil including Mixtures or derivatives thereof, containing synthetically produced or natural Sources are obtained.

additional Objects and attendant advantages of the present invention partially set out in the description that follows, or may be of the To run or using the present invention. The Tasks and benefits can realized and achieved by means and combinations, especially in the attached claims are called. It must be understood that the above general Description and the following detailed description only as an example and explanatory are and not as for the invention as claimed is to be viewed as limiting.

detailed Description of the Preferred Embodiments

It is a pharmaceutical composition for preventing or treating described by breast cancer, the composition being at least a connection of an ethereal Vegetable oil including Mixtures or derivatives of compounds of essential vegetable oils which from either natural or synthetic sources.

The special, here revealed ethereal vegetable oils or derivatives thereof include a monocyclic, carbocyclic Ring structure that is six-membered and with at least one functional Substituted, which is oxidized or comprises hydroxyl. The ethereal vegetable oils the use according to the invention are selected from the group consisting of cinnamaldehyde, benzyl alcohol, cinnamyl alcohol, Carvacrol, citronellal, eugenol, isoeugenol, thymol and trans-anethole. Because these compounds are more ethereal vegetable oils are known and for Other uses can be used by a specialist using known methods.

The The above-mentioned compounds of an essential vegetable oil can be obtained from usual Sources can be related simply isolated and cleaned from special plants or trees (isolated) or can using conventional Techniques are synthesized. Advantageously, these compounds from easily available Starting materials are synthesized. The relative ease with the compositions used in the present invention can be synthesized represents a huge advantage in the production of these compounds in big scale represents.

It will be recognized that the therapeutically active compounds an ethereal one Vegetable oil used in the present invention modified or derivatized by attached suitable functionalities, i.e. functional groups to selective biological properties to reinforce. Such modifications are known in the art and include those who put the biological penetration into a given biological Compartment (e.g., blood, lymphatic system, central nervous system) enlarge, the oral availability enlarge, the solubility enlarge to to allow administration by injection, to change the metabolism and change the excretion rate. In addition, the compounds of an ethereal vegetable oil in pro-drug forms be converted so that the desired therapeutically effective Form of connection in the body of the patient as the result of the action of metabolic or other biochemical processes to the pro-drug is generated. such Examples of prodrug forms include ketal, acetal, oxime and Hydrazone forms of compounds, the ketone or aldehyde groups contain.

In addition, the therapeutically active compounds of an essential vegetable oil for use in the present invention, one or more asymmetric carbon atoms contain and can as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers occur. Each stereogenic carbon may have the R or S configuration. All Such isomeric forms of these compounds are expressly disclosed in U.S. Pat included within the scope of the present invention.

As will be recognized, include the compositions described herein and methods of pharmaceutical compositions containing at least an ethereal one vegetable oil and pharmaceutically acceptable salts thereof, in combination with a any pharmaceutically acceptable carrier, excipient or vehicle include. The term "pharmaceutical acceptable vehicle or excipient " on a carrier or an adjuvant that is given to a patient together with a patient Compound of an essential vegetable oil for use can be administered in the present invention and not the pharmacological activity of which is destroyed and non-toxic, if he is administered in doses sufficient to make a therapeutic Quantity of the connection.

Pharmaceutically acceptable salts of the essential vegetable oil compounds that can be used in this invention include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acid salts include, but are not limited to, acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanopropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate, Hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, Succinate, sulfate, tartrate, thiocyanate, tosylate and undecanoate. Other acids, such as oxalic acid, while they are themselves are not pharmaceutically acceptable, can be used in the preparation of salts used as intermediates in obtaining the compounds used in the invention and their pharmaceutically acceptable acid addition salts.

Salts derived from suitable bases include alkali metal (eg, sodium), alkaline earth metal (eg, magnesium), ammonium, and N- (C _1-4 alkyl) 4 + salts. The present invention also provides for the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil soluble or dispersible products can be obtained by such quaternization.

Further include pharmaceutically acceptable carriers, excipients and vehicles, which can be used in the pharmaceutical compositions without limited to this to be, ion exchanger, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS), such as d.alpha-tocopherol-polyethylene glycol-1000 succinate, or other similar polymeric delivery matrices or systems, serum proteins such as human Serum albumin, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, Glyceride partial mixtures of natural vegetable fatty acids, Water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, Potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, Polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, Sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, Polyethylene glycol and wool fat. Cyclodextrins, such as alpha, beta and gamma-cyclodextrin or chemically modified derivatives, such as Hydroxyalkylcyclodextrins, including 2- and 3-hydroxypropyl-beta-cyclodextrins, or other soluble made derivatives also advantageously used to deliver therapeutic effective compounds of an ethereal Vegetable oil used in the present invention.

The Pharmaceutical compositions can be administered orally, parenterally, by an inhalation spray, topically, rectally, nasally, buccally, vaginally or over implanted reservoir, however, is the oral Administration or injection. The pharmaceutical compositions may be any conventional one non-toxic pharmaceutically acceptable carriers, adjuvants or vehicles contain. In some cases For example, the pH of the formulation may be with pharmaceutically acceptable acids, bases or buffers are adjusted to the stability of the formulated compound or to improve their mode of administration. The term "parenteral" as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injections or infusion techniques.

The Pharmaceutical compositions may be in the form of a sterile injectable preparation, e.g. as a sterile injectable aqueous or fatty suspension available. This suspension can be prepared according to those known in the art Techniques can be formulated using suitable dispersing or wetting agents (such as Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in one non-toxic parenterally-acceptable diluent or solvent be, e.g. a solution in 1,3-butanediol. Among the acceptable carriers and solvents used can be are mannitol, water, wrestlers solution and isotonic sodium chloride solution. additionally are sterile, non-volatile oils conventionally as a solvent or Suspension medium used. For this purpose, any used mild non-volatile oil be inclusive synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives, are in the production of injectable oils, as it is natural pharmaceutical are acceptable oils, like olive oil or castor oil, especially useful in their polyoxyethylated versions. These oil solutions or suspensions can also a diluent or dispersants of a long-chain alcohol, such as Ph.Helv or a similar one Alcohol.

The Pharmaceutical compositions may be in any orally more acceptable Dosage form can be administered, including, but not limited to Capsules, tablets and watery Suspensions and solutions. In the case of tablets for the oral use include carriers, which are commonly used, lactose and cornstarch. Lubricant, such as magnesium stearate, are also typically added. For the oral Administration in a capsule form includes useful diluents Lactose and dried corn starch. When watery Suspensions are administered orally, becomes the effective ingredient combined with emulsifying and suspending agents. If desired, certain Sweet and / or Flavoring and / or coloring matters be added.

The pharmaceutical compositions may also be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and will therefore melt in the rectum to release the active components. Such materials include, but are not limited to, cocoa butter, beeswax, and polyethylene glycols.

Even though Rarely, is the topical administration of the pharmaceutical compositions especially useful, if the desired Treatment surfaces or organs that are easily accessible through topical application are. For An application topically on the skin should be the pharmaceutical composition be formulated with a suitable ointment containing the effective components suspended or dissolved in a carrier contains. carrier for the topical administration of the compounds of this invention but not limited on mineral oil, liquid Mineral oil, White oil, propylene glycol, Polyoxyethylene-polyoxypropylene compound, emulsifier wax and water. Alternatively, the pharmaceutical composition may be treated with a suitable Lotion or cream can be formulated which is the effective compound in a carrier suspended or dissolved contains. Suitable carriers include, but are not limited to on mineral oil, Sorbinsäuremonostearat, Polysorbate 60, wax of cetyl esters, cetearyl alcohol, 2-octyldodecanol, Benzyl alcohol and water. The pharmaceutical compositions can also topically to the lower digestive tract by rectal suppository formulation or in a suitable enema formulation. Topically-transdermal Plasters are also included.

The Pharmaceutical compositions can be delivered by a nasal aerosol or inhalation. Such compositions be according to well known in the art of pharmaceutical formulation techniques made and can as solutions in salt, using benzyl alcohol or other suitable Preservatives, absorption promoters, bioavailability to improve fluorocarbons and / or other solubilizers or dispersants known in the art become.

A other acceptable pharmaceutical preparation would be an encapsulated form the ethereal Vegetable oils, as it is, or modified as described above be. The walls of the Capsules could be designed to be the ethereal vegetable oils release quickly, i. in a minute, hour or day, or they could be designed to over a certain period of time, i. Days, weeks or months to release. The wall materials could natural or synthetic polymers that are acceptable to the US FDA, or consist of lipids or other suitable materials. These capsules could either orally or by injection and could be either on water or oil based, dependent of the desired Method of use or required release rate.

dosage levels from between about 0.001 and about 100 mg / kg body weight per day, preferably between about 0.5 and about 75 mg / kg body weight per day of the compound of the active ingredient are at Prevention and treatment of soft tissue cancers useful. Typically, the pharmaceutical compositions of approximately once to about five times per day or alternatively administered as a continuous infusion become. Such administration may be considered chronic or acute therapy can be used. The amount of active ingredient, the one with the carrier materials can be combined to produce a single dosage form, becomes dependent vary depending on the host treated and the particular mode of administration. A typical preparation is about 5% until about 95% effective compound (w / w) included. Preferred are such preparations of about 20% to about 80% effective compound included.

To Improving the condition of a patient can be a maintenance dose a compound, composition or combination of this invention administered, if necessary. Subsequently, the dosage or frequency of administration, or both, as a function of the symptoms, to a level where the improved state is maintained; if the symptoms are to the desired Level have been alleviated, the treatment should be stopped. Patients can however, a treatment with interruptions on a long-term basis with every recurrence of disease symptoms.

The Prophylactic use of the present invention may be taken daily require a prophylactically effective amount.

As those skilled in the art will appreciate, lower or higher doses than those mentioned above may be required. Special dosage and action regimens for any particular patient will depend on a variety of factors, including the effectiveness of the particular compound used, age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, severity, and the course of a cancer , the patient's disposition towards cancer and the judgment of the attending physician.

The The compositions and methods described herein will be further developed in the following, non-limiting Examples illustrated. The examples are for different embodiments only as an example and restrict not the claimed invention with respect to the materials, Conditions, Weight ratios, Process parameters and the like, which are mentioned here. If the Following these examples, it will be recognized that the growth and proliferation of MCF-7 cells are strictly estrogen-dependent. In the presence of estrogen the cells grow, grow together and form foci, the landmark the tumor diagnosis. In the absence of estrogen, the growth of these is Cells are low and the formation of herds is rare.

example 1

Antiproliferative effect on human epithelial breast cancer cells (MCF-7)

MCF-7 cells were cultured in the growth medium supplemented with 10% fetal bovine serum (FBS). At 85% confluence, cells were subcultured in 5% FBS serum-purified medium, phenol red-free for 24 hours prior to treatment of the test chemical. After 24 hours of treatment, cell proliferation was measured using ³ H-thymidine incorporation. These test chemicals were tested in the presence and absence of estrogen to distinguish whether they have anti-estrogenic activity in addition to their antiproliferative effect. The study was performed in triplicate and a control was used only with solvent. The control received solvent only at <0.1% ethanol. The estrogen was tested at 1 nM (= 0.27 ng E ₂ / ml). Exemplary compounds of an essential vegetable oil were tested at 50 μg / ml. The results are shown in Table 1.

Comparative Example 1

Anti-proliferative effect

MCF-7 cells were cultured in the growth medium supplemented with 10% fetal bovine serum (FBS). At 85% confluence, cells were subcultured in 5% FBS serum-purified medium, phenol red-free for 24 hours prior to treatment of the test chemical. After 24 hours of treatment, cell proliferation was measured using ³ H-thymidine incorporation. These test chemicals were tested in the presence and absence of estrogen to distinguish whether they have anti-estrogenic activity in addition to their antiproliferative effect. The study was performed in triplicate and a control was used only with solvent. The control received solvent only at <0.1% ethanol. The estrogen was tested at 1 nM (= 0.27 ng E ₂ / ml). Compounds of an essential vegetable oil were tested at 50 μg / ml. The results are shown in Table 2.

Example 2

Dose-response effect on E ₂ -induced cell growth

MCF-7 cells were cultured in growth medium containing 10% fetal calf serum (FBS) was supplemented. At 85% confluency, the cells were subcultured in six-well petri dishes and in 5% FBS serum purified medium, phenol red free 24 hours long before the treatment of various concentrations of the test chemicals added. After five Days of treatment, cells were trypsinized and used collected an Eppendorf microcentrifuge. The cell pellets were resuspended in 1% Trypan Blue and three equivalent quantities (10 μl each) the viable Cell suspensions were counted using a hemocytometer assay. Each sample was then counted three times and the data shown is the average of three counts. These test chemicals were in the presence of 10 nM estrogen (= 2.7 ng estrogen / ml) tested. 2 wells per test concentration were used. This Experiment was repeated twice. The control was solvent only at <0.1% Ethanol. The results are shown in Table 3.

Our data show a dose-response relationship of essential vegetable oil compounds and their anti-estrogenicity against E ₂ -induced abnormal cell growth and proliferation in human mammary epithelial cancer cells (MCF-7). These data showed that thymol provides 40% protection against E ₂ -induced abnormal growth in cancerous breast cells at a low dose (5 μg / ml). In addition, these data also showed that eugenol (1 μg / ml) and isoeugenol (1 μg / ml) exhibited 35% and 25% protection, respectively, against E ₂ -induced abnormal growth in cancerous breast cells (see Example 3). Furthermore, cinnamaldehyde provided approximately 96% control of E ₂ -induced abnormal growth in cancerous breast cells at 50 μg / ml (see Example 3).

These Data show, inter alia, that certain essential vegetable oils and combination of it for anti-proliferative, anti-estrogens and / or anti-mitogenic compounds useful for prophylactic or therapeutic treatment of soft tissue cancers are useful.

Even though the illustrative examples of the present invention in Detail have been described, it must be understood that the present Invention is not limited to those precise embodiments, and that different changes and modifications can be effected therein by a person skilled in the art without to depart from the scope of the invention as defined by the appended claims.

Claims (11)

Using a compound of an ethereal vegetable oil selected from eugenol, thymol, isoeugenol, benzyl alcohol, carvacrol, cinnamyl alcohol, Cinnamic aldehyde, citronellal or trans-anethole for the manufacture of a medicament for the treatment or prevention of breast cancer. Use according to claim 1, wherein the compound an ethereal one vegetable oil Eugenol is. Use according to claim 1, wherein the compound an ethereal one vegetable oil Thymol is. Use according to claim 1, wherein the compound an ethereal one vegetable oil Isoeugenol is. Use according to claim 1, wherein the compound an ethereal one vegetable oil Benzyl alcohol is. Use according to claim 1, wherein the compound an ethereal one vegetable oil Carvacrol is. Use according to claim 1, wherein the compound an ethereal one vegetable oil Cinnamon alcohol is. Use according to claim 1, wherein the compound an ethereal one vegetable oil Cinnamon aldehyde is. Use according to claim 1, wherein the compound an ethereal one vegetable oil Citronellal is. Use according to claim 1, wherein the compound an ethereal one vegetable oil trans-anethole is. Use according to claim 1 to 10, wherein the breast cancer E2-induced breast cancer is.